Effects of Bushen Huoxue Decoction on skeletal,vascular,and renal functions in postmenopausal osteoporosis rats based on protein expressions of Klotho,PI3K,and Akt
Objective To observe the effects of Bushen Huoxue Decoction(BSHXD)on the protein expression level of Klotho in the proximal femur,adventitia of aorta,and renal tubules as well as those of phosphatidylinositol 3-kinase(PI3K)and protein kinase B(Akt)in the proximal femur of ovariectomized rats,so as to explore the mechanism of action of BSHXD in treating postmenopausal osteoporosis(PMOP).Methods Thirty-one 6-month-old female SD rats were randomly divided into blank group(n=8),sham-operated group(n=5),and surgery group(n=18).After eight weeks of surgical modeling,three rats from blank group and another three from surgery group were randomly selected and sacrificed.After successful modeling,the remaining rats in the surgery group were subdivided into model group(n=5),BSHXD group(n=5),and estradiol valerate group(n=5)according to the random number table method.Then,blank,sham-operated,and model groups were given distilled water,BSHXD group was given BSHXD 3.32 mg/kg,and estradiol valerate group was given estradiol valerate 0.1 mg/kg.All groups were treated by gavage continuously for eight weeks.The general condition of rats was observed;the serum calcium content was measured by spectrophotometry;the serum phosphorus content was determined by coupled reaction method;bone mineral density of the proximal femur was measured by bone density scanner,and the bone cells were examined by electron microscopy.In addition,the pathological changes in the proximal femur were observed by HE staining;Klotho protein level in the proximal femur,aortic adventitia,and renal tubules was measured by ELISA;the relative expression levels of Klotho,PI3K,and Akt proteins in the proximal femur were checked by Western blot.Results Compared with model group,the rats in BSHXD and estradiol valerate groups showed higher average body weight and better vitality,and there was no significant difference in the condition of diet and hair;the number and form of bone cells and bone tubules in the proximal femur were restored to varying degrees,and the osteoporotic lesions were also repaired to different extent.Compared with blank and sham-operated groups,the serum calcium content of rats in model and BSHXD groups was lower(P<0.01,P<0.05);compared with model group,the serum calcium content of rats in estradiol valerate group was higher(P<0.05).Compared with blank and sham-operated groups,rats in model group showed a significant decrease in the bone mineral density(P<0.01),as well as in the Klotho protein content in the proximal femur,aortic adventitia,and renal tubules(P<0.01);the Klotho protein expression in the bone tissue was significantly reduced(P<0.01),while the protein expressions of PI3K and Akt were significantly elevated(P<0.01).Compared with model group,the Klotho protein content in the proximal femur,aortic adventitia,and renal tubules of rats in BSHXD and estradiol valerate groups increased(P<0.01);the Klotho protein expression in the bone tissue was significantly higher(P<0.01),while the protein expressions of PI3K and Akt were significantly lower(P<0.05,P<0.01).Compared with BSHXD group,the Klotho protein content in the proximal femur and renal tubules of rats in estradiol valerate group increased(P<0.05),while the protein expressions of PI3K and Akt in the bone tissue decreased(P<0.05).Moreover,there was no statistical difference in serum phosphorus content among the groups(P>0.05).Conclusion BSHXD can alleviate the osteoporosis-like lesions of ovariectomized rats to a certain extent by elevating the Klotho protein expression in the proximal femur,aortic adventitia,and renal tubules and inhibiting the protein expressions of PI3K and Akt in the proximal femur.And its mechanism of action may be related to inhibiting osteocyte apoptosis and improving renal function and circulatory system function.
Bushen Huoxue Decoctionpostmenopausal osteoporosisKlotho proteinphosphatidylinositol 3-kinasepro-tein kinase B
国家科技期刊平台
NSTL
万方数据
维普
